CN104610423A - Method for extracting glycoprotein by utilizing trepang processing blanching liquid - Google Patents
Method for extracting glycoprotein by utilizing trepang processing blanching liquid Download PDFInfo
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- CN104610423A CN104610423A CN201510061478.XA CN201510061478A CN104610423A CN 104610423 A CN104610423 A CN 104610423A CN 201510061478 A CN201510061478 A CN 201510061478A CN 104610423 A CN104610423 A CN 104610423A
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- sea cucumber
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- Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
- Y02—TECHNOLOGIES OR APPLICATIONS FOR MITIGATION OR ADAPTATION AGAINST CLIMATE CHANGE
- Y02P—CLIMATE CHANGE MITIGATION TECHNOLOGIES IN THE PRODUCTION OR PROCESSING OF GOODS
- Y02P20/00—Technologies relating to chemical industry
- Y02P20/50—Improvements relating to the production of bulk chemicals
- Y02P20/54—Improvements relating to the production of bulk chemicals using solvents, e.g. supercritical solvents or ionic liquids
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Abstract
The invention discloses a method for extracting glycoprotein by utilizing trepang processing blanching liquid. The method is characterized by comprising the following steps of collecting the glycoprotein blanching liquid and cooling the glycoprotein blanching liquid rapidly to 4-10 DEG C, and removing undissolved substance in the glycoprotein blanching liquid in a centrifugation manner to obtain centrifuged supernate; carying out electrodialysis demineralizing on the centrifuged supernate by adopting an electrical drive device to obtain demineralized clear liquid; concentrating the demineralized clear liquid in a vacuum concentration mode to obtain concentrated liquid with the solid content of 20-45%; freeze-drying or spray-drying the concentrated liquid to obtain a first solid dry material; degreasing the first solid dry material by utilizing petroleum ether or normal hexane or ethanol solution or a supercritical CO2 fluid extraction method to obtain a second solid dry material; adding the second solid dry material into pure water, and heating and extracting to obtain extractive solution; after extracting, removing deposits in the extractive solution by utilizing a centrifugation mode, filtering supernate through a 10Kda membrane, ultrafiltering the filtered solution, and collecting trapped fluid; drying the trapped fluid to obtain the trepang glycoprotein.
Description
Technical field
The present invention relates to a kind of method utilizing Holothurian machining blanching solution to extract glycoprotein.
Background technology
After fishing for, there is the enzyme of proteolysis such as autolytic enzyme in fresh sea cucumber, being easy to skin, self-dissolving due to himself.For the ease of preserving, fresh sea cucumber usually will add and preserves as starting material after hot water scalding treatment.According to incompletely statistics, often process 1 ton of fresh sea cucumber, the sea cucumber blanching solution of 0.6-1.2 ton can be produced.Along with the mass-producing, intensive of Holothurian machining; collectable sea cucumber blanching solution resource is more and more abundanter; how this resource of higher value application; turn waste into wealth; be developed to the functional food with high utility value; extend sea cucumber industrial chain, the problem that the scientific research personnel day by day becoming mass-producing Holothurian machining enterprise and be engaged in sea cucumber research pays close attention to jointly.
A large amount of active substances is contained as selenka (imitating isolation identification journal of Shandong university (Edition) the 7th phase in 2011 of several selenka compound in stichopus japonicus processing-waste), sea cucumber suger albumen (ZL 2,011 1 0384722.8) etc. in sea cucumber blanching solution.Sea cucumber suger albumen has good raising immunizing power, leukocyte increasing quantity, the biological activitys such as adjunct antineoplastic.How more efficiently at present, extract glycoprotein from sea cucumber blanching solution, more the foundation of low cost is applicable to the novel technical method that industrialization is produced, and is an important research direction.
Summary of the invention
Object of the present invention provides a kind of method utilizing Holothurian machining blanching solution to extract glycoprotein.
The technical measures that the present invention takes are: a kind of method utilizing Holothurian machining blanching solution to extract glycoprotein, it is characterized in that it comprises the steps: (1) and collect sea cucumber blanching solution being cooled to 4-10 DEG C rapidly, adopt centrifugation, the insolubles removed in sea cucumber blanching solution obtains centrifuged supernatant;
(2) centrifuged supernatant adopts electrically-actuated device electrodialysis, and desalination, makes conductance reach 300-1000 μ S/cm and stop, obtaining desalination clear liquid;
(3) desalination clear liquid adopts vacuum concentration mode, is concentrated into solid content between 20-45%, obtains concentrated solution;
(4) concentrated solution obtains the first dry solid through lyophilize or spraying dry;
(5) the first dry solid utilized sherwood oil or normal hexane or mass percent for 90-95% ethanolic soln or supercritical CO
2the means such as fluid extraction, carry out skimming treatment, obtain the second dry solid;
(6) join in pure water by the second dry solid, extract solution is extracted to obtain in heating;
(7) after extracting, extract solution removes precipitation through centrifugal Horizontal helical type centrifuge, disc centrifuge, tubular-bowl centrifuge successively, supernatant liquor is by the membrane filtration of 10KDa, and the filter membrane that filtered solution is 5000Da by molecular weight cut-off carries out ultrafiltration, collects trapped fluid.
(8) obtain sea cucumber suger albumen after trapped fluid drying, in this component, protein content is between 80-90% after measured, and sugared content is between 10-20%.
Concrete feature of the present invention also has, and Holothurian machining blanching solution refers to and utilizes the processing blanching solution produced in fresh Holothurian machining blanching process as extraction material.The concrete operation step that Holothurian machining blanching solution obtains is fresh sea cucumber after dissecting and removing intestines, puts into boiling water and boils 10-30min, pulls gained liquid after sea cucumber out.
Beneficial effect of the present invention be remove salinity by electrically-actuated device of lowering the temperature rapidly, utilize, concentrated after, lyophilize or spraying dry obtain dry-matter, effectively ensure that sea cucumber blanching solution fast eliminating salinity when not putrid and deteriorated, utilize sherwood oil or normal hexane or 90-95% ethanolic soln or supercritical CO afterwards
2the compositions such as the lipid in the means effective eliminations such as fluid extraction sea cucumber blanching solution; Recycling membrane technique is separated and obtains the glycoprotein analog component of molecular weight between 5000-10000Da, and this step effectively to eliminate in sea cucumber blanching solution the compositions such as albumen, saponin(e, taurine, small peptide; The combination of each processing step of this novel method, greatly improve the efficiency utilizing sea cucumber blanching solution to extract glycoprotein, greatly reduce ethanolic soln usage quantity, by the method extract obtain sea cucumber suger protein content at protein content between 80-90%, sugar content is between 10-20%, the quantity of leucocyte reduction measured causing after Chemotherapy of Tumor Patients through pharmacological evaluation has significant rising effect, and to quantity of leucocyte, normal patient has no effect.This technological method reduces production cost from the extraction glycoprotein of Holothurian machining blanching solution and equipment investment more, and product has high health care and pharmaceutical use.
Embodiment
embodiment 1:
A kind of method utilizing Holothurian machining blanching solution to extract glycoprotein, it comprises the steps: (1) collects Holothurian machining blanching solution 5000L, be cooled to 4 DEG C rapidly, afterwards successively through horizontal screw centrifuge 3000rpm, disc centrifuge 6000rpm, tubular-bowl centrifuge 12000rpm, after the insolubles in centrifugal segregation sea cucumber blanching solution, collect supernatant liquor.
(2) supernatant liquor adopts electrically-actuated device (the HC-EDR-3 type that Taicang HuaChen Water Treatment Facilities Co., Ltd. produces) to carry out electrodialytic desalting, stops to obtain desalination clear liquid when solution conductivity reaches 600 μ S/cm.
(3) desalination clear liquid utilizes vacuum concentration, and being concentrated into solid content is 30%, obtains concentrated solution;
(4) concentrated solution spray-dried after the first pressed powder.
(5) above-mentioned first pressed powder joins in 600L 95% ethanolic soln, stirs 24h, removes fat, utilize pumping and filtering device suction filtration, collects filter residue, obtains the second pressed powder after filter residue and drying.
(6) second pressed powders join in 2000L pure water, being heated to 100 DEG C keeps 60min to obtain extract solution, extract solution is cooled to rapidly 4 DEG C, first through Horizontal helical type centrifuge (3000rpm), again through disk plate centrifuge (rotating speed 6000rpm), finally by tubular-bowl centrifuge (rotating speed 12000rpm), collected after centrifugation supernatant liquor.
(7) supernatant liquor obtains filtered solution by the membrane filtration of molecular weight cut-off 10KDa, and the filter membrane that filtered solution is 5000Da by molecular weight cut-off carries out ultrafiltration, collects trapped fluid.
(8) trapped fluid drying is sea cucumber suger albumen, after testing protein content 81% afterwards, polysaccharide content 12%.
embodiment 2:
A kind of method utilizing Holothurian machining blanching solution to extract glycoprotein, it comprises the steps: (1) collects Holothurian machining blanching solution 5000L, be cooled to 4 DEG C rapidly, afterwards successively through horizontal screw centrifuge 3000rpm, disc centrifuge 6000rpm, tubular-bowl centrifuge 12000rpm, after removing the insolubles in sea cucumber blanching solution after the insolubles in centrifugal segregation sea cucumber blanching solution, collect supernatant liquor.
(2) supernatant liquor adopts electrically-actuated device to carry out electrodialytic desalting, stops to obtain desalination clear liquid when solution conductivity reaches 500 μ S/cm.
(3) desalination clear liquid utilizes vacuum concentration, and being concentrated into solid content is 35%, obtains concentrated solution.
(4) concentrated solution obtains the first dry solid after vacuum lyophilization.
(5) first dry solid utilize supercritical CO
2fluid extraction (Huaan, Nantong HA420-40-200 type), Extracting temperature is at 50 DEG C, and extractor pressure is at 30Mpa, and extraction 1h, removes fat, must remove the second dry solid of fat.
(6) above-mentioned the second dry solid removing fat joins in 2000L pure water, being heated to 100 DEG C keeps 60min to obtain extract solution, extract solution cools rapidly latter 4 DEG C, first through Horizontal helical type centrifuge (3000rpm), again through disk plate centrifuge (rotating speed 6000rpm), finally by tubular-bowl centrifuge (rotating speed 12000rpm), collected after centrifugation supernatant liquor.
(7) supernatant liquor is by the membrane filtration of molecular weight cut-off 10KDa, and the filter membrane that filtered solution is 5000Da by molecular weight cut-off carries out ultrafiltration, collects trapped fluid.
(8) trapped fluid drying is sea cucumber suger albumen, after testing protein content 83% afterwards, polysaccharide content 13%.
Claims (2)
1. utilize Holothurian machining blanching solution to extract the method for glycoprotein, it is characterized in that it comprises the steps: (1) and collect sea cucumber blanching solution being cooled to 4-10 DEG C rapidly, adopt centrifugation, the insolubles removed in sea cucumber blanching solution obtains centrifuged supernatant;
(2) centrifuged supernatant adopts electrically-actuated device electrodialysis, and desalination, makes conductance reach 300-1000 μ S/cm and stop, obtaining desalination clear liquid;
(3) desalination clear liquid adopts vacuum concentration mode, is concentrated into solid content between 20-45%, obtains concentrated solution;
(4) concentrated solution obtains the first dry solid through lyophilize or spraying dry;
(5) the first dry solid utilized sherwood oil or normal hexane or mass percent for 90-95% ethanolic soln or supercritical CO
2fluid extraction means, carry out skimming treatment, obtain the second dry solid;
(6) join in pure water by the second dry solid, extract solution is extracted to obtain in heating;
(7) after extracting, extract solution removes precipitation through centrifuge mode, and supernatant liquor is by the membrane filtration of 10KDa, and the filter membrane that filtered solution is 5000Da by molecular weight cut-off carries out ultrafiltration, collects trapped fluid;
(8) sea cucumber suger albumen is obtained after trapped fluid drying.
2. the method utilizing Holothurian machining blanching solution to extract glycoprotein according to claim 1, it is characterized in that the centrifuge mode in step (7) is successively through centrifugal Horizontal helical type centrifuge by extract solution, disc centrifuge, precipitation removed by tubular-bowl centrifuge.
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Cited By (1)
Publication number | Priority date | Publication date | Assignee | Title |
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CN105029513A (en) * | 2015-08-20 | 2015-11-11 | 大连润伟海参研发科技有限公司 | Method for extracting saponin, polysaccharide, peptide and melanin in wastewater obtained by boiling sea cucumbers |
Citations (3)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
CN102406048A (en) * | 2011-11-29 | 2012-04-11 | 山东好当家海洋发展股份有限公司 | Method for preparing sea cucumber glycoprotein by using sea cucumber blanching solution |
CN102657320A (en) * | 2012-04-16 | 2012-09-12 | 浙江工业大学 | Cryo-concentration method of juice obtained by processing organisms |
CN103725740A (en) * | 2013-12-06 | 2014-04-16 | 山东好当家海洋发展股份有限公司 | Method for extracting glycoprotein by using scollop blanching liquid |
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Patent Citations (3)
Publication number | Priority date | Publication date | Assignee | Title |
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CN102406048A (en) * | 2011-11-29 | 2012-04-11 | 山东好当家海洋发展股份有限公司 | Method for preparing sea cucumber glycoprotein by using sea cucumber blanching solution |
CN102657320A (en) * | 2012-04-16 | 2012-09-12 | 浙江工业大学 | Cryo-concentration method of juice obtained by processing organisms |
CN103725740A (en) * | 2013-12-06 | 2014-04-16 | 山东好当家海洋发展股份有限公司 | Method for extracting glycoprotein by using scollop blanching liquid |
Cited By (1)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
CN105029513A (en) * | 2015-08-20 | 2015-11-11 | 大连润伟海参研发科技有限公司 | Method for extracting saponin, polysaccharide, peptide and melanin in wastewater obtained by boiling sea cucumbers |
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